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Related Experiment Videos

Enzyme-labelled immunoassay for plasma cortisol.

T Ogihara, K Miyal, K Nishi

    The Journal of Clinical Endocrinology and Metabolism
    |January 1, 1977
    PubMed
    Summary

    A new enzyme-labelled immunoassay for plasma cortisol offers a reliable method for clinical analysis. This technique accurately measures cortisol levels, showing good correlation with radioimmunoassay results.

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    Area of Science:

    • Biochemistry
    • Immunology
    • Clinical Chemistry

    Background:

    • Plasma cortisol measurement is crucial for diagnosing various endocrine disorders.
    • Existing methods like radioimmunoassay may have limitations in routine clinical settings.

    Purpose of the Study:

    • To develop and validate a novel enzyme-labelled immunoassay (ELISA) for accurate plasma cortisol quantification.
    • To assess the suitability of this new assay for routine clinical laboratory use.

    Main Methods:

    • Cortisol was conjugated to alkaline phosphatase via 21-hemisuccinate using water-soluble carbodiimide.
    • Antibodies were generated against cortisol-21-hemisuccinate-bovine serum albumin in rabbits.
    • Plasma samples were pre-treated by boiling to denature endogenous proteins, followed by a double antibody method for separation of bound and free cortisol.

    Main Results:

    • The developed enzyme-labelled immunoassay demonstrated a linear relationship between plasma volume and cortisol concentration.
    • The assay achieved a minimal detectable level of 1 mug/dl for plasma cortisol.
    • High precision was observed with coefficients of variation of 2.7%-4.4% (within-assay) and 4.7%-6.0% (between-assay).
    • Results showed excellent correlation with established radioimmunoassay methods.

    Conclusions:

    • The enzyme-labelled immunoassay provides a sensitive, precise, and reliable method for plasma cortisol determination.
    • This assay is well-suited for routine clinical analysis, offering an alternative to radioimmunoassay.

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